Synchrotron radiation as a light source in confocal microscopy of biological processes

Hans C. Gerritsen; C. J. R. van der Oord; Yehudi K. Levine; Ian H. Munro; Wendy J. Myring; D. A. Shaw; Fokko F.G. Rommerts

doi:10.1117/12.58271

1 April 1992 Synchrotron radiation as a light source in confocal microscopy of biological processes

Hans C. Gerritsen, C. J. R. van der Oord, Yehudi K. Levine, Ian H. Munro, Wendy J. Myring, D. A. Shaw, Fokko F.G. Rommerts

Author Affiliations +

Proceedings Volume 1640, Time-Resolved Laser Spectroscopy in Biochemistry III; (1992) https://doi.org/10.1117/12.58271
Event: OE/LASE '92, 1992, Los Angeles, CA, United States

Abstract

A novel confocal microscope is presented using the Daresbury Synchrotron Radiation source as its light source. The broad spectrum of synchrotron radiation in combination with the UV compatible microscope allows the extension of confocal microscopy from the visible to the UV region down to about 200 nm. It is envisaged that structures separated by about 70 nm can be resolved at a wavelength of 200 nm. In addition, the tunability of synchrotron radiation affords the selective excitation of any specific fluorescent molecule at the maximum of the absorption band. This avoids the restriction of working at fixed laser lines. A further advantage of using synchrotron radiation is the realization of multiwavelength excitation. Test results using laser systems in the visible and in the UV are presented. Fluorescence images of test targets using UV excitation reveal the superior resolution of the microscope. Furthermore, images of Leydig cells incubated with a fluorescent cholesterol derivative whose maximum of absorption is at 325 nm are shown. These images cannot be produced by conventional confocal laser microscopes. Finally, promising preliminary results obtained with synchrotron radiation are presented.

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Hans C. Gerritsen, C. J. R. van der Oord, Yehudi K. Levine, Ian H. Munro, Wendy J. Myring, D. A. Shaw, and Fokko F.G. Rommerts "Synchrotron radiation as a light source in confocal microscopy of biological processes", Proc. SPIE 1640, Time-Resolved Laser Spectroscopy in Biochemistry III, (1 April 1992); https://doi.org/10.1117/12.58271

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